绿色荧光蛋白gfp基因在钝顶螺旋藻A9藻株中的表达
Expression of Green Fluorescent Protein Gene in A9 Strain of Spirulina platensis
将钝顶螺旋藻（Spirulina platensis）A9藻株在24℃培养，经2mmol/L EDTA预处理24h；采用功率300w的超声波处理70s获得单细胞样品，以本实验室构建的携带gfp基因的质粒p215t转化A9藻株单细胞藻液，利用amp作为选择标记，使单细胞在平板上再生长出单藻落，获得17株具有Amp抗性的转化藻株，转化率3.73‰。在390nm紫光激发下，生长30d的转化藻丝体发出稳定绿色荧光；培养45d后具有绿色荧光的藻丝出现断裂、具有荧光藻丝的长度缩短的现象。试验结果初步表明报告基因gfp在螺旋藻中得到稳定有效的表达，可以采用单细胞再生形成单藻落技术进行螺旋藻的基因克隆。
9 strain of Spirulina platensis was cultured at 24℃ and treated with 2 mmol/L EDTA for 24 hours. Single cell system was obtained by ultrasonic treatment with power of 300w for 70s. The sample of single cell system was transformed with plasmid p215t with gfp gene, and amp was used as selective marker. The single cell sample was successfully regenerated as single colonies on plates with Amp and 17 transformants were obtained with transformation rate being about 3.73‰. Observed by fluorescence microscope with 390nm as stimulation light, parts of the transforments filaments presented stable green fluorescence when grew for 30d. After 45d part of the green fluorescent transformants trichomes began to break and the length of the trichomes with green fluorescent were shortened. The results showed that techniques of the single cell regeneration could be applied for gene cloning in Spirulina, and the foreign gfp gene has been effectively expressed in S. platensis.
张明、魏娟、张海生、唐欣昀、叶霁、曹媛媛
分子生物学微生物学生物工程学
钝顶螺旋藻 gfp基因 转化 单细胞再生 绿色荧光
Spirulina platensis gfp gene transformation single cell regeneration green fluorescence
张明,魏娟,张海生,唐欣昀,叶霁,曹媛媛.绿色荧光蛋白gfp基因在钝顶螺旋藻A9藻株中的表达[EB/OL].(2007-04-11)[2025-05-04].http://www.paper.edu.cn/releasepaper/content/200704-274.点此复制
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